delta cholenates and process for preparing same



Unite States Patent 2,854,462 A CHOLENATES ANDSPROCESS FOR PREPARING Earl M. Chamberlin, Westfield, and John M Chemerda,

Metuchen, N. J., assignors to Merck & Co., Inc., Rahway, N. J a corporation of.-New Jersey No Drawing. Original applications March 10, 1951, Se-

rial No. 215,026, and September 20, 1951, Serial No. 247,562. Divided and .this application November 26, 1957, Serial No. 703,721

2, Claims. c1. 260-3911 This application is a division of copending applications Serial No. 215,026, filed March 10, 195'1,now abandoned, and Serial No. 247,562, filed September 20, 1951.

This invention is concerned with novel chemical compounds of the cyclopentanopolyhydrophenanthrene series and processes for preparing the same; more particularly, it relates to novel cyclopentanopolyhydrophenanthrene compounds having functional substituents in ring and specificallyit relates to'new compounds having ahydrox-yl or 'keto substituent at the '11 position, andto processes for thepreparation of such compounds.

Compounds of the adrenal cortex, such as 'Kendalls Compound E (cortisone), have been foundto be of great value in the treatment of various diseases. Further, it is likely that Kendalls Compound ;E and/or other closely related ll-hydroxy steroids Will find increasing therapeutic use in the future. Unfortunately, the only method for the preparation of such compounds presently available utilizes desoxycholic or cholic acids as *thetstarting material. Cholic and desoxycholic acids have Ihydroxy substituents in ring 'C at the 12 position, .thus providinga means for introducing :a functional 'substituent at-the 11 position. However cholic and desoxycholic acids, which are obtained :from animal bile, :are only available in limited amounts. Heretofore :no practical method ;was available whereby:a;functional :groupcould be introduced in ring Which'would permit the use of more abundant steroids such as the sterols, ergosterol, :cholesterol, stigmasterdl, or plant sapogenins, such :as :diosgenin, itigogenin, 'andrthe 311126.

It is an object of the apresentiinvention to provide a processfor introducingafunctional group in rin'g C:at the 11 position. It is a further object to provide a process for converting cyclopentanopolyhydrophenanthrene compounds having a double bond in the 7:8 position to the corresponding -cyclopentano1;iolyhydrophenanthrene "compoundhavin g a hydroxyl or keto group at positions )7 :and 11. Another object is to provide new compounds o'f-the steroid .scrieshaving tfunctional groupsfin ringtC suitable for .the ,pr.epar ation of other ;cyclopentan opolyhydro-' phenanthrene compounds. ,Other objects will beiapparent from [the detailed-description hereinafter provided.

.In .accordance withour invention, we {have now found that compounds of the -cyclopentanopolyhydrophenanthrene .series having :an Il-ketosubstituent-can betsynthesizedby reactions :indicatedos follows 2,854,462 Patented Sept. 30, 1958 VII These reactions are carried out as follows:

qyclqp msnopolyhyd p snaathr ne qmpoundthaving-a ,7 ii-double bond (1;) is reacted with mercuric acetate producing the corresponding "compound having conjue ted double bonds in the 7:8 .a1i 19:11 positions (=II) wh c zi t ea ed-W t a-pe 'aci t s-f rm g ncp x de repre e e y t al ernat ormu a 11A and III- (At present the exact structure ofthe epo'xide is not-known and it is represented by th e alternative :forms 1111A :and H113. Alternatively :it ;is possible that :the product ,obtained is a -mixture :of these two forms.) This epoxide is then treated with an adsorbent to form the corresponding -,A -.J, -.1-dihyd oxy compou The h fl ll dihydroxy compound 1(IV) isreactedwith an oxidizing agent to convert the "hydroxy substituents to lgeto groups, :thus obtaining the corresponding A -7,11- dike'to :cyclopentanopolyhydrophenanthrene derivatives (-,V). These diketo compounds are then reduced to saturatethegkbdouble bondand'forrn the corresponding 7,11- diketo compound VI. The saturated diketo-compound is thenred'uceduto eliminate the 'l keto substituent,-thus producing thetcorresponding ll-keto compound (VII); lattercompoundsare useful intermediates-for the preparation .of :lil-keto .compounds having desirable therapeutic prop erties. The 1-A l '12;compounds of the cyclopentanopolyhydrophenanthrene series are conveniently prepared by react ingsthe correspondingxiv-compound with mercuric acetate. We have found that this reaction'is preferablyeliected by reacting :the A -compound with mercuric acetate and glacial acetic acid in thepresence of a suitable solvent medium such as chloroform. T he reaction is conveniently conducted by stirring the reaction mixture for 16-24 hours. After the reaction'is completed, the A -compound is recovered fro n the reaction mixture by removing-theprecipitated mercurous acetate, and concentrating the solution under diminished pressure. If desired, the residue may be further purified =by crystallization from snitable solvents. Thus, this process can be utilized to prepare .iAW kpregnadienerii-o120-one-3acetate, and A a -dehydrotigogenin acetate from .A'Epre'gnenolone 3 acetate and A -dehydrotigogenin acetate respectively. Alternatively, other acyl derivatives of these starting materials or the 3-hydroxy compounds may be utilized as starting materials in our process to prepare the corresponding A' -compounds. t

Further, the 3-hydroxy-A" -choladienic acid, which is also useful as a starting material in the processes of our invention, is readily obtained by reducing 3-hydroxy 12-keto-A -choladienic acid. Thisis conveniently accomplished by reacting the keto acid with'hydrazine hydrate and an alkali metal hydroxide in the presence of a suitable high boiling solvent such as diethylene glycol.

This invention is concerned with compounds of theLtype represented by intermediates V and VI above, and with processes of producing the same. Compounds V and V1 7 may be represented by the following formulas:

The starting materials used in the process of this invention, namely the A -dihydroxy compounds shown by Formula IV, may beobtained as described in copending application Serial No. 215,026, now abandoned, filed March 10, 1951.

Pursuant to our invention, we have found that the a,

A-7,1l-dihydroxy compounds are readily oxidized to obtain a new series of compounds having a double bond in the 8:9 position and keto substituents in the 7 and 11 positions. The oxidation is readily accomplished for example by treating the A-7,1l-dihydroxy compounds with chromioacid-in' the presence of an acid such as acetic or sulfuric acid. We have found that an oxidation mixture consisting of chromic acid-acetone-sulfuric acid is particularly useful for this oxidation, and results in the ob-' tainment of maximum yields of the desired diketo' compound under optimum conditions. In carrying out this process, it is necessary to protect any other hydroxyl substituents, for example, a 3-hydroxy group, by converting thisgroup to an acyloxy substituent. After the oxidation, any such acyloxy substituentscan be readily hydrolyzed to prepare the corresponding hydroxy compounds.

These new A'-7,11-diketocyclopentanopolyhydrophenanthrene compounds are reduced to saturate the 8:9 double bonds and produce the corresponding saturated 7,11- diketo; compound. This reductionis conveniently accomplished for example by reacting the A compound with zinc in the presence of acetic acid.

In accordance with the above-described methods, cyclopentanopolyhydrophenanthrene. compounds having 7,11- diketo substituents such as, 3-acyloxy-7,1l-diketo-A' ergostadiene, 3-acyloxy-7,1 1-diketo-A bisnorallocholenic acid and its esters, 3-acyloxy-7,ll-diketorA cholenic acid and its esters, 3-acyloxy-7,1lrdiketo-A -tigogenin, A'-allopregnene-3-ol-7,ll 20rtrione' and its acyl. derivatives, and the like can be obtained by the oxidation of the-corresponding 7,1l-dihydroxycompounds. Further, these diketo compounds may be reduced to thecorresponding compounds having a saturated bond in the 8:9 position. 7,

The following examples .are presented to illustrate specific embodiments of our invention.

EXAMPLE 1;

Preparation of 3-acetoxy-7,I1-diketb-A-?'-ergostadlene from 3-acetoxy-7,I I-dihydr0xy-A g ergostadiene Analynla-Calc. for G l-L 0 C, 76.88; H, 9.46.

Found: C, 76.91; H, 9.58.

Max. 2660 A. (E% 200iso-octane) Max.'2700 A. (E% 186ethanol) a +18i2 (1% CHC],)

EXAMPLE 2 Preparation of 3-acet0xy-7,1I-diketo-A""-ergostadiene from 3-acet0xy-7,1 I -dihydr0xy-A -ergostadiene .-To a stirred suspension of 2.36 g. of 3-acetoxy-7,11-dihydroxy-A -ergostadiene in 50 ml. of purified acetone was added'a solution of 6.65 .millimols of chromic oxide in 5 ml. of 3.6 N sulfuric acid. The mixture was stirred at 10. C.; for 10 minutes, after which 2 ml. water was added, and themixture was stirred for 25 minutes at room temperature. The green inorganic material which had separatedHwas removed by filtration. The filtrate was stirred while50 ml. water were added. The product was collected on a filter and washed with water. After drying in vacuo over phosphorous pentoxide the product, S-acetoxy-7,11-diketo-A '"-ergostadiene, was obtained in the form of alight yellow powder. Yield: 2.26 mg., M. P. l03-ll4fC. max. 2660 (E% l21ethanol).

EXAMPLE 3 Preparation of--3-acetoxy-7,Il-diketo-A -ergostene from '3-acetoxy-7,I1-diket0-A""-ergostadiene A hot solution of m. of 3-acetoxy-7,1l-diketo-A' ergostadiene in'5 cc. of acetic acid and 0.1 cc. of water was treated with 250 mg. of zinc. The initially yellow solution was decolorized rapidly and the mixture heated on the steam bath for three hours. Water was then added to precipitate the diketone, which was dissolved in benzene. Upon removal of the benzene, a residue was ob tained which was recrystallized from methanol. Yield: 70 mg; M. P. 196-198 C.; a --25 (1.0% CHCI,).

Analysis.'Calc. for G l-1 ,0 C, 76.55; H, 9.85. Found: ,C, 76.68; H, 9.59. Alkaline hydrolysis of 3-acct oxy-7,ll-diketo-A -ergostene yielded 3-hydroxy-7,ll-diketo-A -ergostene, M. P. 198-199 C.

Analysis.Calc. for C l-L 0 C, 78.46; H, 10.37. Found: C,.78.28; H, 10.09.

EXAMPLE 4 Preparation of methyl 3-acetoxy-7,lI-diketo-A'-blsnoral- I, locholenate from methyl 3-acetoxy-7JI-dihydroxy- A'-bisn0rall0cholenate To a' stirred solution 'of'1.60 g. of methyl 3-acetoxy- 7,11-dihydroxy-A -bisnorallocholenate in 30 ml. of glacial acetic acid, was added a solution of 0.730 g. of sodium dichromate dihydrate in '30 ml. of glacial acetic acid over a period of 15 minutes. After stirring at 25 C. for two two recrystallizations from methanol, the melting point of the diketo compound was raised to'186.5-187.5' C. [a] "=+38.7 (C=1.04, CHCl,); A max. 2700; E% 212.

*5 Analysis.-Calc. for o gn o r C, 69.74; H, 7.96. Found: C, 70.00; H, 8.12. V w

EXAMPLES. Methyl 3-hydr0xy-1J keto-bisnorallocholanate O-O'OH H it VIII LIV One gram of 3 a'cetoxy 7;-1 1+dlketo-1S er'gostene (I) obtained as described in Example 3 was dissolved in 100 cc. of chloroform and ozone was pa's'se'd' throug'h =at=ice bath temperature until the approximate -'theoretical amount of ozone was absorbed. "The reaction mixture was diluted with 100 cc. of -'glaci-'al acetic *acid, c'ooled "to 5 C. and oxidized with'llfj 'g.'chromic=acid dis'sdlved 'in 0.75 cc. water and 50 cc. glacial acetic ac'id.

After standing overnight, 5 cc. of methanol was added and the solvent was removedin-vacuo to practically dryness. The residue in the flask was dissolved by shaking twice with a mixture of 25 cc. of 5% sulfuric acid and 50 cc. of benzene. The combined benzene solutions were dried over anhydrous magnesium sulfate, and the benzene was evaporated on the stream bath in a stream of nitrogen.

The residue was dissolved in 200 cc. of ether and stirred with 5 g. of sodium carbonate and 2 cc. of water for 21 hours. The sodium salt of '3-acetoxy-7,1l-diketobisnorallocholanic acid (II) was filtered off and dried in a vacuum desiccator.

The dried sodium salt was suspended in 25 cc. of ether and 25 cc. of 50% sulfuric acid was added in small portions until the mixture was definitely acid. 100 cc. of ether was added to bring about complete solution of all solids. The aqueous layer was separated and extracted once with 50 cc. of ether.

The combined ethereal solutions were dried over anhydrous magnesium sulfate and then evaporated on the steam bath to a small volume, whereupon 3-acetoxy-7,lldiketo-bisnorallochloanic acid (II) crystallized out. The product was recrystallized from ether; M. P. 235-238 C. [cc] =24.6, a=0.68; C=l.38% CHCl Analysis.Calcd for C H O C, 68.87; H, 8.19. Found: C, 68.67; H, 8.04.

125 mg. of 3-acetoxy-7,l1-diketo-bisnorallocholanic acid was suspended in 25 cc. of ether and esterified with diazomethane. All solid dissolved and on evaporation of the ether to a small volume, the methyl ester (III) crystallized. M. P. 226.5-229 C. Mixed melting point with an authentic sample of the ester: 227230 C.

5 g. of methyl 3-acetoxy-7,ll-diketo-bisnorallocholanate (III) and 2.07 g. of powdered potassium hydroxide were placed in a 50 cc. round-bottom flask. 25 cc. of

diethylene glycol rand.2.3 cc. offi% hydrazine hydrate were added and the temperature raised to -140 -C. andlheld for lhour. The temperature was then raised to 200 C. and heId for-.2 hours.

After, cooling, the reaction'm'ixture was dissolved in benzene and water, 50% sulfuric acid .added until an acid reaction was obtained. The benzene layer was separated, and the aqueous layer extracted three times with 50 cc. 'of benzene. The tc'ombined benzene solutions were washed with water anddr ied over anhydrous nnagnesium sulfate.

The benzene was treated with Darco. The "benzene soldtionwa's 'concentratedinyacuo 'to "dryness, the' residue was -dissolvedfin ether "and 'esterified with 'an ethereal solution of diazjome'thane. "The ether was evaporated, and the methyl B-hydroxy-"Il-keto "bisnorallocholanate ((IV') was recrystallized from methanol, P. 1775-- 1805. Mixed with'an authenticsample, 177-179" "C. [a i-41.

EXAMPLE6 Preparation pf methyl 3-.a'cetoxy-7,1lediketa fisnvralio holan'ate from methyl 3-zacet0xy- -Z'1l-fiket0 A lrisnorallocholenate A mixture of 400 g. of methyl 3-acetoxy-7,1l-diketo- A -bisnorallocholenate, 20ml. of g1acial acetic acid, 0.4 ml. water, .and 1.5 .g. .zinc dust washeated for one hour on the steam bath. One gram of zinc .dust was added, and the mixture was heated for two hours more. After coolingto room tempera'ture, 50 ml. water were added to the mixture. The precipitated product, along with res'idual' zinc, was collected on a filter. The dried mixture was extracted with benzene. After removal of the *ben zene by vacuum distillation, theproduct, methyl S-acetoxy-"7,'1"-l diketo bisnorallocholanate was recrystallize'd from "methanol. Yield: 270 mg. P. 230-2515 C. The analytical sample wasrecrystallized from methanol, MLPS-2'30i5-23L5 C. 1061 -l-'4:5 (C= 1.55 {CI-R31 a lnalysia- Galc. for 6 1 1 0 C, '69541;H, 8.39. Found: C, 69.70; 51,840. e

EXAMPLE 7 Preparation of methyl 3-acetoxy-7J1-diketo-A -cholenate from methyl 3-acet0xy-7,1I-dihydroxy-M-cholenatc To a stirred suspension of 1.7 g. of methyl 3-acetoxy- 7,1l-dihydroxy-A -cholenate in 100 cc. acetone was added at 20 C. with stirring a solution of 0.5 g. chromium trioxide in 4 cc. of 10% sulfuric acid over a period of ten minutes. After addition was complete, the mixture was stirred for one and one half hours. The inorganic residue was filtered ofi and washed with three 10 cc. portions of acetone. The acetone solution was added with stirring to 66 cc. water and the 7,11-diketo compound which precipitated was filtered, washed free of acid with water and dried. Yield: 1.57 g.; M. P. 112.5-- 113.5 C.

After recrystallization from ethanol the product, methyl 3-acetoxy-7,1l-diketo-n -cholenate melted at 114-115 C.

Analysis.-Calc. for C H O C, 70.40; H, 8.75. Found: C, 70.93; H, 8.30.

EXAMPLE 8 Preparation of methyl 3-acetoxy-7,1 11aliketa-ch0lwnme from methyl 3-acet0xy-7,1I-diketo-M-cholenate A mixture of 1.57 g. of methyl 3-acetoxy-7,11-diketo- A -cholenate, 17 cc. acetic acid, 1.7 cc. water and 3.4 g. zinc dust was heated on a steam bath for one hour.

The reaction mixture was diluted with 80cc. benzene and the zinc residue was filtered oil. The benzene solution was washed free of acid with water, dried over 7 anhydrous sodium sulfate and concentrated to dryness. Yield l.57g. I I

The product was purified by recrystallization from ethanol. Yield: 1.17 g.; M; P-. 162-3 C. V Analysis-Cale. for CflH O C, 70.10; H, 9.15. Found: C, 70.52; H, 8.65. "L

.. X M L l 7 Preparation of 3-acetoxy-7,ll-diketo-8 dchydrotigbgenih from 3-acetoxy-7JIdihydrbxy-8-dehydrotigogenin' To a suspension of 0.346g. (0.71.mil1imol) of 3- acetoxy-7,ll-dihydroxy-8-dehydrotigogenin in 10 m1. of acetone was added 0.8 ml. of a solutionvof 1.06 millimols of chromium trioxide in 3.6 N sulfuric acid. After stirring for 15 minutes, the chromium salts which had formed were removed by filtration and werewashed with ml. of acetone. The filtrate was .treated with :50-60 ml. of water to precipitate the product which, after drying,

weighed 0.283 g., M. P, l90 -20 0 C. After two recrystallizations from methanol, 3-acetoxy-7,ll-dikcto-8-dehydrotigogenin was-obtained as pale yellow needles, M. P. 226-227 0. th, -14 --(C.=0.813', CHCl,) x max. (in ethanol) 2700,

EXAMPLE 10 Preparation of 3-acet0.t:y-7,II-eiiketotigogenin from 3 acetoxy-7,II-diketo-8-dehydrotigogenin A mixture of 130 mg. of 3-acetoxy- 7,1l-diketo-S-dehydrotigogenin, 4 ml. glacial acetic acid, one drop of water, and 400 mg. of zinc dust was heated on the steam bath .for one hour. The mixture was cooled, and 30 ml. of water and-'20 ml. of chloroform were added. After shaking thoroughly and filtering, the layers were separated, and the aqueous layer was extracted twicevwith five ml. portions of chloroform. TIhe combined chloroformcxtracts were dried over sodium sulfate, and concentrated in vacuo. Recrystallization of the residue gave small rectangular prisms. Yield 75 mg., M. P. 24l-243 C., lai 72 (c.=0.8 27, CHCl,).

EXAMPLE 12 Preparation of 'aIloprcgmnJ-ol-ZII,20-tri0nc-3-acctate from M-alIopregnene-J-ol-ZI] ,20-trione-3-acetate A mixture of 210 mg. of A' -allopregnene-3-ol- 7,11,20-trione-3-acetate, 10 ml. of glacial acetic acid, two drops of water, and 1.0 g. of zinc dust was heated on the steam bath for 1 hour. The product was isolated in the usual manner, and recrystallized from methanol to give mg. of hexagonal prisms, M. P. 214-215 C.

Various changes and modifications may be made in carrying out the present invention without departing from the spirit and scope thereof. Insofar as these changes and modifications are within the purview of the annexed claims, they are to be considered as part of our invention.

We claim:

1. Lower alkyl' esters of 3-acyloxy-7,ll-diketo-A'- cholenic acid wherein the acyl substituent is a lower fatty acidradical, V

,2. Methyl 3-acetoxy-7,1l-diketo-N-cholenate.

3. A process for the conversion of methyl A'-3-acetoxy- 7,l-1-dihydroxy-cholenate into thecorresponding methyl A'-3-.ac etoxy 5 7,11 diketo cholenate, which comprises treating the methyl A'-3-acetoxy-7,l1,dihydroxy-cholenate with chromic. acid.

. ,No references cited. 

1. LOWER ALKYL ESTERS OF 3-ACYLOXY-7,11-DIKETO-$8CHOLENIC ACID WHEREIN THE ACYL SUBSTITUENT IS A LOWER FATTY ACID RADICAL. 